Construction and identification of chromokinesin KIF4A knockdown gastric cancer cell lines
10.3760/cma.j.issn.1673-4181.2014.01.005
- VernacularTitle:染色体驱动蛋白KIF4A稳定低表达胃癌细胞系的构建及鉴定
- Author:
Kaize ZHONG
;
Jian ZHAO
;
Pei LI
;
Weiwen CHEN
;
Changjun ZHU
;
Zhixiong DONG
- Publication Type:Journal Article
- Keywords:
KIF4A;
Mitosis;
Spindle midzone;
siRNA;
Gastric cancer cell
- From:
International Journal of Biomedical Engineering
2014;37(1):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct stable chromokinesin KIF4A knockdown gastric cancer cell lines (SGC-7901) and study the mitotic spindle midzone formation in these cells.Methods Short hairpin RNA (shRNA) expression plasmids of pGPU-GFP-shKIF4A,pGPU-GFP-shNC specific targetting KIF4A and non-sense DNA sequences were constructed respectively and then transfected into SGC-7901 cells.The cells were cultured in selection medium containing G418 to form single clones.Stable KIF4A shRNA expressing SGC-7901 cells were picked up under an fluoroscent microscope and identified by Western Blot.The spindle midzone in these cells was analyzed after immunofluorescence staining.Results Three cell lines stably expressing KIF4A shRNA and one with shNC were successfully constructed.Compare to the control cell line,KIF4A knockdown cell lines represented remarkable elongation of spindle midzone and the less the KIF4A,the longer spindle midzone.Conclusions Stable KIF4A knockdown SGC-7901 cell lines were successfully constructed that can serve for further study of KIF4A ' s function and its role in proliferation and development of gastric cancer.
